Evaluation of acrosomal integrity of frozen-thawed simmental bull sperm using different staining methods

Fertility rates that vary depending on the male animal whose semen is used in artificial insemination or natural breeding are of great importance. For this reason, it is of great importance to know the potential fertility of male animals in order to provide information about fertility and sterility. Sperm motility and kinematic evaluations are powerful parameters that help predict the fertility ability of spermatozoa. However, these examinations do not give full information about the fertility ability. The motility of the spermatozoon and its active movement in the linear direction provide information about the ability to pass through the female reproductive tract. A successful acrosome reaction is important for fertilization. Activation of the acrosome at the right time, release of enzymes, and facilitating penetration of spermatozoa into the zona pellucida demonstrate the importance of acrosome integrity for successful fertilization. Acrosome integrity plays a crucial role in fertility success. A wide variety of staining procedures are used in the evaluation of acrosome integrity. These procedures are mostly used to determine the percentage of defective spermatozoa and acrosomes. In this direction, in our thesis project, it is aimed to compare and evaluate the acrosome integrity in frozen thawed Simmental bull semen using different staining methods. This study showed that Trypan Blue Giemsa (TBG), Spermac and Coomassie Blue staining is a simple, inexpensive and reliable staining method for the evaluation of acrosome integrity in Simmental bull semen. With these methods, spermatozoa can be evaluated quickly by performing both morphological examination and dead-alive examination in a single smear. Considering that these staining methods are very easy and can be easily applied in field conditions, it can be said that this method is a very advantageous method, considering the morphological characteristics of spermatozoa very quickly. When these three staining methods were compared, Coomassie Blue staining showed the highest positive correlation with spermatological parameters. As a result, both Trypan blue-Giemsa and Spermac methods, especially Coomassie blue, are simple, inexpensive and effective methods that work fast in frozen-thawed semen of Simmental bulls, do not require complex laboratory conditions in the calculation of dead viviparous and acrosomal/morphological disorders, and are quickly used in routine analyses and should take its place.